NF-kappaB-independent induction of endothelial cell apoptosis by the vascular disrupting agent DMXAA.

DMXAA (5,6-dimethylxanthenone-4-acetic acid; AS1404), a vascular disrupting agent currently in clinical trials, induces tumour endothelial cell apoptosis in vivo in mice and in cancer patients. DMXAA activates NF-kappaB in many different cell types. In this study, whether DMXAA-induced endothelial cell apoptosis was NF-kappaB dependent was determined.HUVEC endothelial and T24 endothelial-like cells were treated with DMXAA and apoptosis was measured by terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL). NF-kappaB activation was measured by electrophoretic mobility shift assays (EMSA). T24 cells were transfected with IkappaBalphaM, a mutant form of the IkappaBalpha gene which cannot be phosphorylated and degraded, hence preventing NF-kappaB expression.No NF-kappaB up-regulation was detected in apoptotic HUVEC treated with DMXAA. The IkappaBalphaM-transfected T24 cells showed similar apoptotic responses to those of parental cells.The DMXAA-induced apoptosis is neither mediated by, nor inhibited by, the expression of the NF-kappaB pathway.